Sep 19, 2023
Protocol for "Neuromelanin accumulation drives endogenous synuclienopathy in non-human primates"
- jlanciego1,2,3
- 1CNS Gene Therapy Program, Center for Applied Medical Research (CIMA) University of Navarra;
- 2Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CiberNed-ISCIII);
- 3Aligning Science Across Parkinson's (ASAP) Collaborative Research Network
Collection Citation: jlanciego 2023. Protocol for "Neuromelanin accumulation drives endogenous synuclienopathy in non-human primates". protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6xdwrlqe/v1
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this collection and it's working
Created: September 19, 2023
Last Modified: September 19, 2023
Collection Integer ID: 87996
Keywords: Tyrosinase, Alpha-synuclein, Lewy bodies, Marinesco bodies, Prion-like spread
Funders Acknowledgement:
Aligning Science Across Parkinsons (ASAP)
Grant ID: ASAP-020505
Spanish Ministry of Science and Innovation
Grant ID: PID2020-120308RB-I00
CiberNed Intramural Collaborative Projects
Grant ID: PI2020/09
Disclaimer
The authors report no competing interests
Abstract
This study was aimed to develop and characterize
a non-human primate (NHP) model of Parkinson’s disease mimicking the known
neuropathological hallmarks of Parkinson’s disease to the best possible extent.
Accordingly, we sought to determine whether AAV-mediated enhanced expression of
human tyrosinase (hTyr) in the substantia nigra (SNpc) of non-human primates
(NHPs) is able to induce a time-dependent accumulation of neuromelanin (NMel)
in dopaminergic neurons, further triggering and endogenous synucleinopathy,
progressive cell death and a pro-inflammatory scenario, in keeping with what
was formerly reported in rats by taking advantage of a similar strategy (Carballo-Carbajal
et al., 2019). Furthermore, the potential prionoid spread of endogenous
alpha-synuclein (a-Syn) species towards the
prefrontal cortex was analyzed, in an attempt to evaluate to what extent there
is a propagation of endogenous a-Syn
by permissive trans-synaptic templating (e.g. the so-called Braak hypothesis).
Adult juvenile NHPs (Macaca fascicularis)
were injected with adeno-associated viral vectors (AAVs) encoding either the
hTyr gene (AAV-hTyr; delivered into the left SNpc) or a null construct for
control purposes (AAV-null; injected into the right SNpc). In order to
delineate a timeline for the underlying processes, one group of NHPs was
sacrificed four months post-AAV deliveries (animals M308F4 and M310M4), whereby
the follow-up timing for second experimental group was settled at eight months
post-AAVs surgeries (animals M307F8 and M309M8). Neuroimage studies (MRI and
MicroPET) were conducted in vivo at
different time points. Upon animal sacrifices, brain tissue samples were
processed for histological analysis comprising intracellular NMel levels,
intracellular aggregates, nigrostriatal degeneration and neuroinflammation.
Attachments
PROTOCOL.docx
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Protocol references
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